Involvement of Rac and Rho signaling in cancer cell motility in 3D substrates.

The motility of cancer cells in 3D matrices is of two types: mesenchymal motility, in which the cells are elongated and amoeboid motility, in which the cells are round. Amoeboid motility is driven by an actomyosin-based contractile force, which is regulated by the Rho/ROCK pathway. However, the molecular mechanisms underlying the motility of elongated cells remain unknown. Here, we show that the motility of elongated cells is regulated by Rac signaling through the WAVE2/Arp2/3-dependent formation of elongated pseudopodia and cell-substrate adhesion in 3D substrates. The involvement of Rac signaling in cell motility was different in cell lines that displayed an elongated morphology in 3D substrates. In U87MG glioblastoma cells, most of which exhibit mesenchymal motility, inhibition of Rac signaling blocked the invasion of these cells in 3D substrates. In HT1080 fibrosarcoma cells, which display mixed cell motility involving both elongated and rounded cells, inhibition of Rac1 signaling not only blocked mesenchymal motility but also caused a mesenchymal-amoeboid transition. Additionally, Rac1 and RhoA signaling regulated the mesenchymal and amoeboid motility in these cells, respectively, and the inhibition of both pathways dramatically decreased cell invasion. Hence, we could conclude that Rac1 and RhoA signaling simultaneously regulate cell invasion in 3D matrices.

Diabetes mellitus prevents ischemic preconditioning in patients with a first acute anterior wall myocardial infarction.

OBJECTIVES: This study was undertaken to assess whether prodromal angina could have beneficial effects in diabetic patients with acute myocardial infarction (AMI). BACKGROUND: Prodromal angina occurring shortly before the onset of AMI is associated with favorable outcomes by the mechanism of ischemic preconditioning. However, little is known about the impact of diabetes on ischemic preconditioning. METHODS: We studied 611 patients with a first anterior wall AMI who underwent emergency catheterization within 12 h after the onset of chest pain: 490 patients without diabetes and 121 patients with non-insulin treated diabetes. Prodromal angina was defined as angina episode(s) occurring within 24 h before the onset of AMI. Serial contrast left ventriculograms were obtained in 424 patients at the time of acute and predischarge catheterization. RESULTS: In non-diabetic patients, prodromal angina was associated with lower peak creatine kinase (CK) value (3,068 +/- 2,647 IU/l vs. 3,601 +/- 2,462 IU/l, p = 0.037), larger increase in left ventricular ejection fraction (LVEF) (10.1 +/- 13.0% vs. 5.8 +/- 13.4%, p = 0.004) and lower in-hospital mortality (3.4% vs. 9.3%, p = 0.015). On the contrary, in diabetic patients, there was no significant difference in peak CK value (3,382 +/- 2,520 IU/l vs. 3,233 +/- 2,412 IU/l, p = NS), the change in LVEF (6.7 +/- 13.8% vs. 7.1 +/- 12.4%, p = NS) and in-hospital mortality (8.8% vs. 11.0%, p = NS) between patients with and patients without prodromal angina. CONCLUSIONS: Prodromal angina limited infarct size, enhanced recovery of LV function and improved survival in non-diabetic patients with AMI. However, such beneficial effects of prodromal angina were not observed in diabetic patients, suggesting that diabetes might prevent ischemic preconditioning.

Impact of diabetes mellitus on long term survival after acute myocardial infarction in patients with single vessel disease.

OBJECTIVE: To assess the influence of diabetes on long term prognosis after reperfusion treatment and its interaction with multivessel disease. DESIGN: A retrospective observational study. SETTING: Hiroshima City Hospital. PATIENTS: 1660 consecutive patients with acute myocardial infarction who underwent coronary angiography within 24 hours after the onset of chest pain. MAIN OUTCOME MEASURES: Influence of diabetes on 10 year survival after infarction was assessed using the generalised Wilcoxon test and Cox's proportional hazards regression. Follow up was completed in 1622 patients (98%). RESULTS: Diabetic patients had more multivessel disease than non-diabetic patients (53% v 34%, p < 0.001). When only patients with single vessel disease were compared, diabetes was associated with a reduced 10 year survival after infarction (p = 0.002). On the other hand, in patients with multivessel disease there was no significant difference in survival between diabetic and non-diabetic patients (p = 0.70). Multivariate analysis also showed that diabetes was an independent risk factor related to 10 year mortality after infarction in patients with single vessel disease (odds ratio (OR) 1.81, 95% confidence interval (CI) 1.27 to 2.54; p = 0.001) and not in patients with multivessel disease (OR 1.17, 95% CI 0.85 to 1.60; p = 0.34). CONCLUSIONS: Diabetes is an independent predictor of long term mortality after infarction in patients with single vessel disease. However, in the presence of multivessel disease, prognosis after infarction is impaired regardless of diabetes, and the influence of diabetes is less obvious.

Apoptosis is not increased in myocardium overexpressing type 2 angiotensin II receptor in transgenic mice.

To determine whether angiotensin type 2 (AT(2)) receptor stimulation induces apoptosis in cardiomyocytes in vivo, we developed transgenic mice overexpressing the AT(2) receptor in a cardiac-specific manner, using the alpha-myosin heavy-chain promoter. Ten- to 12-week-old male homozygous transgenic mice (n=44) and wild-type mice (n=44) were used. Both transgenic and wild-type mice were given either saline (control), a subpressor dose of angiotensin II (100 ng. kg(-1). min(-1)), a pressor dose of angiotensin II (1000 ng. kg(-1). min(-1)) for 14 days, a pressor dose of angiotensin II for 28 days to investigate the effects of stimulation on both angiotensin type 1 (AT(1)) and AT(2) receptors, the AT(1) antagonist L158809 alone, or a combination of angiotensin II (1000 ng. kg(-1). min(-1)) and L158809 for 14 days to investigate the effects of selective AT(2) receptor stimulation. Apoptosis was analyzed in paraffin-embedded ventricular sections by the terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling (TUNEL) technique. In both transgenic and wild-type mice, administration of a subpressor dose of angiotensin II, L158809, or a combination of angiotensin II and L158809 did not significantly affect the tail-cuff blood pressure or heart-to-body weight ratio, whereas administration of a pressor dose of angiotensin II for 14 or 28 days significantly increased blood pressure and the heart-to-body weight ratio. However, there was no statistical difference between the effects of angiotensin II in transgenic and wild-type mice. The number of TUNEL-positive nuclei was approximately 0 to 10 per 100 000 cardiomyocytes, with no difference between transgenic and wild-type mice, regardless of saline infusion or any stimulation. In infarcted canine myocardial tissue sections for positive control, the number of TUNEL-positive nuclei was increased by 13.8 to 19.1 times compared with those in the noninfarcted myocardium. In conclusion, angiotensin II infusion for a period of 28 days failed to induce cardiomyocyte apoptosis regardless of the presence or absence of cardiac AT(2) receptor overexpression. It is unlikely that in mice the AT(2) receptor is a strong signal to induce cardiomyocyte apoptosis in vivo.

Quantitation of DNA damage by an aldehyde reactive probe (ARP).

DNA is constantly damaged by endogenous and environmental agents. Abasic sites representing a major class of DNA damage can be quantitated by an ELISA-like assay using an aldehyde reactive probe (ARP). It is shown that oxidative pyrimidine lesions can be also quantitated by the ARP assay in combination with the treatment with endonuclease III.

[Continuous hepatic arterial infusion of 5-fluorouracil with leucovorin for unresectable liver metastases from colorectal cancer].

Twenty-three patients with liver metastases from colorectal cancer were treated by continuous hepatic arterial infusion chemotherapy with 5-FU and Leucovorin. The regimen was that 500 mg/body of 5-FU with 30 mg/body of Leucovorin was administered continuously for 5 days, followed by no medication for 16 days. The effect of this therapy was evaluated and the relationship between this therapy and the overexpression of vascular endothelial growth factor (VEGF) or microvessel density (MVD) was also studied. Complete response was obtained in 4 patients and partial response in 3 patients; the response rate was 32%. The response rate was 60% in patients who underwent more than 7 courses. The response rate was 44% in patients with positive VEGF and 33% in patients with negative VEGF. The response rate was 50% in patients with an MVD of more than 30 and 33% in patients with an MVD of less than 30. The 3-year survival rate for patients who underwent more than 7 courses was 37.5%. This therapy had to be abandoned in 6 patients due to occlusion of the catheter. Skillful maintenance of the catheter is necessary for a high response rate and satisfactory prognosis using this therapy.

[Long-term prognosis after reperfusion therapy for acute myocardial infarction in elderly patients].

Although it has been well demonstrated that TIMI grade 3 flow is associated with improved survival after acute myocardial infarction in non-elderly patients, its implication in elderly patients has not been clarified. To assess this issue, 1,115 patients with acute myocardial infarction who underwent coronary angiography within 24 hours after the onset of chest pain were studied: there were 131 elderly patients (age > or = 75 years) and 984 non-elderly patients (age < 75 years). Follow-up was achieved for 1,092 patients (98%). Elderly patients were associated with more female, Killip class > or = 2, 3 vessel disease and non-smokers. Although modality of reperfusion therapy was not different, final TIMI flow grade was less frequently obtained in elderly patients (53% vs 65%, p = 0.005). Elderly patients were associated with higher in-hospital mortality (25% vs 9%, p < 0.001) and lower 10 years cardiac death free rate (p < 0.001). Cox proportional hazards model showed that final TIMI flow grade 3 was an independent predictor of 10 years cardiac death free in elderly patients (odds ratio (OR) = 0.39, 95% confidence interval (CI) = 0.20-0.74, p = 0.004) as well as non-elderly patients (OR = 0.41, 95% CI = 0.29-0.58, p < 0.001). In conclusion, our data suggest that final TIMI grade 3 flow is an important determinant to improve short- and long-term survival after acute myocardial infarction in elderly patients as well as in non-elderly patients.

[A case of pulmonary actinomycosis radiologically mimicking a lung cancer].

A 59-year-old male clerk consulted in general practitioner due to cough and hemoptysis. A mass shadow was pointed out in the left upper lung field on a chest radiograph. Patient was referred to our hospital for further treatment. Any definitive daiagnosis could not be made after examinations including sputum culture, cytology and TBLB. Because a lung cancer was strongly suspected, an exploratory thoracotomy was performed. Actinomyces was detected by pathological study of excised specimen, with no evidence of cancer. ABPC was administered for two months postoperatively. The patient is doing well without recurrence of actinomycosis 2.5 years after the surgery. Pulmonary actinomycosis presenting a mass shadow on a radiograph may mimick a pulmonary tumor, especially a lung cancer. Pulmonary actinomycosis should be considered in a differential diagnosis of pulmonary lesion thought to be malignant.

A comparison of angiotensin-converting enzyme inhibitors, calcium antagonists, beta-blockers and diuretic agents on reactive hyperemia in patients with essential hypertension: a multicenter study.

OBJECTIVES: The purpose of this study was to compare the effect of different antihypertensive agents, calcium antagonists, angiotensin-converting enzyme (ACE) inhibitors, beta-blockers and diuretic agents on endothelial function. BACKGROUND: Endothelial dysfunction is a component of essential hypertension, and various antihypertensive drugs may be able to restore normal function. METHODS: Forearm blood flow (FBF) was measured in 296 patients with essential hypertension, including 46 untreated subjects using strain-gauge plethysmography during reactive hyperemia and after sublingual administration of nitroglycerin (NTG). Forty-seven normotensive subjects were similarly evaluated as control subjects. RESULTS: The FBF during reactive hyperemia in the 296 hypertensive patients was significantly less than that in age-matched normotensive subjects. The increase in FBF after administration of sublingual NTG was similar in both groups. Systolic and diastolic blood pressures and forearm vascular resistance were greater in the untreated group than in the four treated groups and did not differ with respect to the antihypertensive agent used. The maximal FBF response from reactive hyperemia was significantly greater in the ACE inhibitor-treated group than in the group treated with calcium antagonists, beta-blockers, diuretic agents, or nothing (40.5 +/- 5.2 vs. 32.9 +/- 5.8, 34.0 +/- 5.6, 32.1 +/- 5.9, and 31.9 +/- 5.8 ml/min per 100 ml tissue, p < 0.05, respectively). Reactive hyperemia was similar in the calcium antagonist, beta-blocker, diuretic and untreated groups, and changes in FBF after sublingual NTG administration were similar in all groups. The infusion of NG-monomethyl-L-arginine, a nitric oxide (NO) synthase inhibitor, abolished the enhancement of reactive hyperemia in hypertensive patients treated with ACE inhibitors. CONCLUSIONS: These findings suggest that ACE inhibitors augment reactive hyperemia, an index of endothelium-dependent vasorelaxation, in patients with essential hypertension. This augmentation may be due to increases in NO.

Abnormal magnesium status in patients with cardiovascular diseases.

To investigate magnesium status in patients with cardiovascular diseases and in those presenting high factors for these diseases, we measured the concentrations of serum total Mg, serum ionized Mg and intra-erythrocyte Mg. Mg is an important cofactor for many enzymes, especially those involved in phosphate transfer reactions. Mg deficiency has been shown to be associated with fatal cardiovascular diseases, as well as with risk factors for these diseases. Only measurement of the serum concentration of total Mg is routinely available, but ionized Mg is the physiologically active component. Furthermore, most of the body's Mg is present in the intracellular space. Subjects included patients with ischaemic heart disease (n=80), cardiac arrhythmia (n=60), diabetes mellitus (n=36), essential hypertension (n=194) and hypercholesterolaemia (n=60). The same measurements were made in healthy controls (30 men and 26 women; mean age 58+/-11 years). The serum ionized Mg concentration was measured with a selective ion electrode. The intra-erythrocyte Mg concentration was measured by atomic absorption. No gender difference was found for any Mg parameter, nor was age related to any Mg parameter. The serum albumin concentration was positively correlated only with the serum total Mg concentration. Although the serum total Mg concentration was similar in all groups, patients with diabetes mellitus and arrhythmia had lower serum levels of ionized Mg. Patients with essential hypertension exhibited higher intra-erythrocyte Mg concentrations than the healthy controls. Thus the measurement of serum total Mg concentration may obscure the presence of extracellular Mg deficiency in patients with arrhythmia and diabetes mellitus. Furthermore, the intracellular accumulation of Mg does not support the hypothesis of Mg deficiency in patients with essential hypertension.

Traumatic cardiac herniation diagnosed by echocardiography and chest CT scanning: report of a case.

A 62-year-old man who had suffered fractures of the left chest wall and hemothorax 1 year earlier was admitted to our hospital with multiple injuries as a result of a traffic accident. Chest drainage was immediately performed under the diagnosis of left hemothorax followed by immobilization of fractures of the femur and radius. On the second day after admission, he suddenly developed severe chest pain and dyspnea after sitting up in bed. Echocardiography and chest computed tomography (CT) showed herniation of the left ventricle into the left thoracic cavity, whereby a diagnosis of cardiac herniation due to rupture of the pericardium was made and an emergency surgical repair was successfully performed. Only eight cases of patients surviving after traumatic cardiac herniation have been reported in Japan; however, this is the first in which a preoperative diagnosis was made by echocardiography and chest CT scanning prior to surgical repair.

Regular aerobic exercise augments endothelium-dependent vascular relaxation in normotensive as well as hypertensive subjects: role of endothelium-derived nitric oxide.

BACKGROUND: Several nonpharmacological interventions, including exercise, are recommended in primary prevention of hypertension and other cardiovascular diseases in which the pathogenetic role of endothelial dysfunction has been suggested. We studied the effects of long-term aerobic exercise on endothelial function in patients with essential hypertension. METHODS AND RESULTS: The forearm blood flow was measured by strain-gauge plethysmography. The responses of forearm vasculature to acetylcholine were smaller in the hypertensive patients than in the normotensive subjects. There was no significant difference in forearm vascular responses to isosorbide dinitrate in the normotensive and hypertensive subjects. We evaluated the effects of physical exercise for 12 weeks on forearm hemodynamics in untreated patients with mild essential hypertension who were divided randomly into an exercise group (n=10) and a control group (n=7). After 12 weeks, the forearm blood flow response to acetylcholine increased significantly, from 25.8+/-9.8 to 32.3+/-11.2 mL. min(-1). 100 mL tissue(-1) (P<0.05), in the exercise group but not in the control group. The increase in the forearm blood flow after isosorbide dinitrate was similar before and after 12 weeks of follow-up in both groups. The infusion of N(G)-monomethyl-L-arginine abolished the exercise-induced enhancement of forearm vasorelaxation evoked by acetylcholine in the exercising group. In normotensive subjects also, long-term aerobic exercise augmented acetylcholine-stimulated nitric oxide release. CONCLUSIONS: These findings suggest that long-term physical exercise improves endothelium-dependent vasorelaxation through an increase in the release of nitric oxide in normotensive as well as hypertensive subjects.

Vasodilating effect of benidipine hydrochloride in the renal and hindquarter vascular regions (supplied by terminal aorta) of spontaneously hypertensive rats.

One of two Ca antagonists, benidipine (3-30 microg/kg) or nifedipine (30-600 g/kg), was administered in a bolus injection through the jugular vein, and the changes in mean arterial pressure (MAP), renal flow (RF), and hindquarter flow (HQF) in conscious spontaneously hypertensive rats (SHRs) and normotensive control rats (NCRs). Renal vascular resistance (RR) and hindquarter resistance (HQR) were calculated as MAP divided by RF and HQF, respectively. When a high dose was administered to decrease the blood pressure by about 20%, the RR was significantly lower with benidipine than with nifedipine. The decrease in HQR was not significantly different between benidipine and nifedipine. When a low dose was administered to decrease the blood pressure by about 7%, the decrease in RR was not significantly different between benidipine and nifedipine, but the HQR was significantly lower with benidipine than with nifedipine. In the NCRs, no pharmacological properties were significantly different between these two Ca antagonists.

Daily aerobic exercise improves reactive hyperemia in patients with essential hypertension.

The effects of long-term aerobic exercise on endothelial function in patients with essential hypertension remain unclear. To determine whether endothelial function relating to forearm hemodynamics in these patients differs from normotensive subjects and whether endothelial function can be modified by continued physical exercise, we randomized patients with essential hypertension into a group that engaged in 30 minutes of brisk walking 5 to 7 times weekly for 12 weeks (n=20) or a group that underwent no activity modifications (control group, n=7). Forearm blood flow was measured using strain-gauge plethysmography during reactive hyperemia to test for endothelium-dependent vasodilation and after sublingual nitroglycerin administration to test endothelium-independent vasodilation. Forearm blood flow in hypertensive patients during reactive hyperemia was significantly less than that in normotensive subjects (n=17). Increases in forearm blood flow after nitroglycerin were similar between hypertensive and normotensive subjects. Exercise lowered mean blood pressure from 115.7+/-5.3 to 110.2+/-5.1 mm Hg (P<0.01) and forearm vascular resistance from 25.6+/-3.2 to 23. 2+/-2.8 mm Hg/mL per minute per 100 mL tissue (P<0.01); no change occurred in controls. Basal forearm blood flow, body weight, and heart rate did not differ with exercise. After 12 weeks of exercise, maximal forearm blood flow response during reactive hyperemia increased significantly, from 38.4+/-4.6 to 47.1+/-4.9 mL/min per 100 mL tissue (P<0.05); this increase was not seen in controls. Changes in forearm blood flow after sublingual nitroglycerin administration were similar before and after 12 weeks of exercise. Intra-arterial infusion of the nitric oxide synthase inhibitor NG-monomethyl-L-arginine abolished the enhancement of reactive hyperemia induced by 12 weeks of exercise. These findings suggest that through increased release of nitric oxide, continued physical exercise alleviates impairment of reactive hyperemia in patients with essential hypertension.

Laparoscopic orchiopexy of intraabdominal undescended testis associated with congenital diaphragmatic hernia.

Laparoscopy is an advantageous method for the repair of intraabdominal undescended testis since both an abdominal exploration and vascular elongation can be effectively performed by laparoscopic assistance. A 3-year-old boy and a 1-year-old boy complaining of unilateral nonpalpable left testes were observed following previous congenital diaphragmatic hernia repairs on the day of birth. In these operations, a laparoscopic working sheath was inserted through a small supraumbilical incision. On each boy, a laparoscopically intraabdominal testis was found close to the left internal inguinal ring. In addition, a 10-mm trocar port was placed in the right lower abdominal quadrant and a 5-mm port was placed in the left lower quadrant. For the purpose of orchiopexy, the left testicular vessels were isolated over their full length. The left side of the testis was pulled through the inguinal ring to an inguinal canal divided by the abdominal wall, and then was retracted into the scrotum. The internal ring was thereafter closed externally by the fascia transversalis. Both patients are doing well, with no testicular atrophy or inguinal hernia for 1 and 2 years, respectively, after the above operations.

[Lung laceration caused by penetrating chest wall injury in Hanshin & Awaji earthquake: a case report].

A 36-year-old man having a left lower lobe laceration caused by penetrating chest wall injury was operated on 5 hours after Hanshin & Awaji Earthquake. At thoracotomy, significant destruction of the left lower lobe was observed. Therefore, we gave up repairing a lung, and performed left lower lobectomy, with a satisfactory outcome. Our hospital is located on 30 km south of seismic center, but the function of our hospital was not completely paralyzed. In Awaji island, emergent treatments were done satisfactorily.